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. 2020 Nov 6;10:19256. doi: 10.1038/s41598-020-75779-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effects of blood constituent incubation with T47D cells—controls. TMED media-treated T47D cells, TMEDWB WB co-incubated with media-treated T47D cells, TMEDPRP PRP co-incubated with media-treated T47D cells, TMEDPPP PPP co-incubated with media-treated T47D cells, TDIL diluent-treated T47D cells, TDILWB WB co-incubated with diluent-treated T47D cells, TDILPRP—PRP co-incubated with diluent-treated T47D cells, TDILPPP PPP co-incubated with diluent-treated T47D. (a) Platelet ultrastructural alterations. White *—membrane folds, black *—platelet hyalomere spread, white arrow—extending filipodia, black arrow—microparticles, white circle—fibrin, black circle—fibrin deposits. A: TMEDWB—slightly active platelet with filipodia and microparticles; B: TDILWB—mostly resting platelets; C: TMEDPRP—spread platelet, filipodia and microparticles. D: TDILPRP—platelets with smooth membrane and extending filipodia, with microparticles. E: TMEDPPP: fully spread platelets with fibrin deposits. F: TDILPPP: fibrin plaque formation. (b) Co-localisation of ERα (green) and ERβ (red) in T47D cells, following exposure to blood constituents. A: TMED: primarily cytoplasmic ERα (white arrow) and ERβ expression in the nucleus (*). B: TMEDWB: ERα and ERβ expression in the nucleus (*). C: TMEDPRP: cytoplasmic and nuclear ERα expression (arrow) and nuclear ERβ (*). D: TMEDPPP: high cytoplasmic ERα (white arrow) and nuclear ERβ (*). E: TDIL—diffuse cytoplasmic and nuclear ERα (white arrow), nuclear ERβ (*). F: TDILWB—cytoplasmic and nuclear ERα (white arrow), higher nuclear ERβ (*). G: TDILPRP—cytoplasmic and nuclear ERα and nuclear ERβ (*). H: TDILPPP—high nuclear and cytoplasmic ERα (black *) and higher nuclear ERβ (white *). (c) Box and whisker plots representing quantitative analysis of ERα (green) and ERβ (red) expression in T47D cells, following exposure to blood constituents. A: Conditioned media-treated T47D cells. B: Diluent-treated T47D cells (0.1% DMSO). ^[Significant differences between ERα and ERβ within the treatment group. *Significant (p < 0.05) differences between matched ERs compared to untreated T47D.